PRODUCT CODE: ET7110-11

Phospho-KAP1 (S824) Recombinant Rabbit Monoclonal Antibody [JE50-99] (ET7110-11)

  • Recombinant

Applications

  • WB

  • IHC-P

  • IP

  • Dot-blot

REACTIVITY

  • Human

Western blot analysis of KAP1 (phospho S824) on 293 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-11, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of KAP1 (phospho S824) on 293 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-11, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-KAP1 (phospho S824) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-11, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-KAP1 (phospho S824) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-11, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue using anti-KAP1 (phospho S824) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-11, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Western blot analysis of KAP1 (phospho S824) on 293 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-11, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • IHC-P

  • IP

  • Dot-blot

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Phospho-KAP1 (S824) Recombinant Rabbit Monoclonal Antibody [JE50-99] (ET7110-11)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding ser824 of human kap1.

Host

Rabbit

Modification

Phospho

Modification Site

S824

Positive Control

293 cell, human tonsil tissue, human breast tissue, human gastric carcinoma tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JE50-99

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

88/79/117 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:2,000

  • IHC-P

  • 1:50-1:200

  • IP:1:10-1:50

TARGET

UNIPROT #

PROTEIN NAME

Phospho-KAP1 (S824)

SYNONYMS

E3 SUMO protein ligase TRIM28 antibody; E3 SUMO-protein ligase TRIM28 antibody; FLJ29029 antibody; KAP 1 antibody; KAP-1 antibody; KRAB associated protein 1 antibody; KRAB interacting protein 1 antibody; KRAB-associated protein 1 antibody; KRAB-interacting protein 1 antibody; KRIP 1 antibody; KRIP-1 antibody; KRIP1 antibody; Nuclear corepressor KAP 1 antibody; Nuclear corepressor KAP-1 antibody; RING finger protein 96 antibody; RNF96 antibody; TF1B antibody; TIF1 beta antibody; TIF1-beta antibody; TIF1B antibody; TIF1B_HUMAN antibody; Transcription intermediary factor 1 beta antibody; Transcription intermediary factor 1-beta antibody; Trim28 antibody; Tripartite motif containing 28 antibody; tripartite motif containing protein 28 antibody; Tripartite motif-containing protein 28 antibody

SEQUENCE SIMILARITIES

Belongs to the TRIM/RBCC family.

TISSUE SPECIFICITY

Expressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.

POST-TRANSLATIONAL MODIFICATION

ATM-induced phosphorylation on Ser-824 represses sumoylation leading to the de-repression of expression of a subset of genes involved in cell cycle control and apoptosis in response to genotoxic stress. Dephosphorylation by the phosphatases, PPP1CA and PP1CB forms, allows sumoylation and expression of TRIM28 target genes.; Sumoylation/desumoylation events regulate TRIM28-mediated transcriptional repression. Sumoylation is required for interaction with CHD3 and SETDB1 and the corepressor activity. Represses and is repressed by Ser-824 phosphorylation. Enhances the TRIM28 corepressor activity, inhibiting transcriptional activity of a number of genes including GADD45A and CDKN1A/p21. Lys-554, Lys-779 and Lys-804 are the major sites of sumoylation. In response to Dox-induced DNA damage, enhanced phosphorylation on Ser-824 prevents sumoylation and allows de-repression of CDKN1A/p21.; Auto-ubiquitinated; enhanced by MAGEA2 and MAGEC2.; Citrullinated by PADI4.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

TIF1β is a member of the TIF1 (transcriptional intermediary factor 1) family, a group of transcriptional regulators that play key roles in development and differentiation. Members of this family are characterized by the presence of two conserved motifs – an N-terminal RING-B box-coiled-coil motif and a C-terminal PHD finger and bromodomain unit. TIF1β is a corepressor for KRAB (Kruppel associated box) domain containing zinc finger proteins. The KRAB domain containing zinc finger proteins are a large group of transcription factors that are vertebrate-specific, varied in their expression patterns between species, and thought to regulate gene transcription programs that control speciation. TIF1β has been shown to be essential for early embryonic development and spermatogenesis. It functions to either activate or repress transcription in response to environmental or developmental signals by chromatin remodeling and histone modification. The recruitment and association of TIF1β with heterochromatin protein (HP1) is essential for transcriptional repression, and for progression through differentiation of F9 embryonic carcinoma cells. TIF1β also plays a role in the DNA damage response. Phosphorylation of TIF1β on Ser842 occurs in an ATM-dependent manner in response to genotoxic stress and is thought to be essential for chromatin relaxation, which is in turn required for the DNA damage response.